Production of beta-glucosidase, hemicellulase and ligninase in E1 and FLC-cellulase-transgenic plants

a technology of hemicellulase and ligninase, which is applied in the field of transgenic plants, can solve the problems of harm to plant growth and development, and the heterologous e1 enzyme does not have a direct access, so as to increase the amount of hydrolyzing enzyme and increase the biomass

Inactive Publication Date: 2007-08-16
BOARD OF TRUSTEES OPERATING MICHIGAN STATE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0020] Therefore, the present invention provides a transgenic plant capable of expressing one or more cell wall degrading enzymes comprising: at least one DNA comprising a cell wall degrading enzyme coding region operably linked to a nucleotide sequence encoding a signal peptide directing the cell wall degrading enzyme encoded by the DNA to an apoplast, plastid or vacuole of the transgenic plant; and at least one DNA comprising a flowering

Problems solved by technology

The third concern was whether increasing the level of production of these heterologous enzymes within the plant cells would cause harm to plant growth and development.
First, heterologous E1 enzyme does not have a direct access to t

Method used

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  • Production of beta-glucosidase, hemicellulase and ligninase in E1 and FLC-cellulase-transgenic plants
  • Production of beta-glucosidase, hemicellulase and ligninase in E1 and FLC-cellulase-transgenic plants
  • Production of beta-glucosidase, hemicellulase and ligninase in E1 and FLC-cellulase-transgenic plants

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0114] This example shows the construction of plasmids comprising a heterologous gene expression cassette comprising a DNA encoding a cellulase fusion protein and a heterologous gene expression cassette comprising a DNA encoding the bar gene (Table 1).

TABLE 1ConstructPlasmid features1rbcSP / e1 / pin 3′ / / Act1rbcSP leaf-specificP / bar / nos 3′promoter drivingcellulase cDNA of A. cellulolyticus2rbcSP / cbh1 / pin 3′ / / Act1rbcSP leaf-specificP / bar / nos 3′promoter drivingcellulase cDNA of T. reesi3rbcSP / rbcS SP / e1 / pin 3′ / / Act1The rbcS SP targetsP / bar / nos 3′cellulase of A. cellulolyticusintomaize chloroplasts4rbcSP / rbcS SP / cbh1 / pin 3′ / / The rbcS SP targetsAct1 P / bar / nos 3′cellulase of T. reesiinto maize chloroplasts

Abbreviations:

The term “rbcSP” means the rice rubisco rbcS promoter region. The rbcSP is a leaf-specific promoter that limits transcription of rbcS to the leaves (Schaeffer and Sheen, Plant Cell 3: 997-1012 (1991)). The nucleotide sequence for the rbcS promoter region is set forth in SEQ...

example 2

[0131] This example shows the construction of plasmids comprising a heterologous gene expression cassette comprising a DNA encoding a cellulase fusion protein. The plasmid constructs are shown in Table 2.

TABLE 2ConstructPlasmid features1rbcSP / cbh1 / pin 3′rbcSP leaf-specificpromoter drivingcellulase cDNA of T. reesei2rbcSP / rbcS SP / cbh1 / pin 3′The rbcS SP targetscellulase of T. reesiinto maize chloroplasts3rbcSP / rbcS SP / syn-cbh1 / pin 3′The rbcS SP targetsmodified cellulase of T. reeseiinto maizechloroplasts4CaMv35s / SSU / e1 / nos3′The SSU targets thecellulase of A. cellulolyticusintomaize chloroplasts5CaMv35s / VSP / e1 / nos3′The VSP targets thecellulase of A. cellulolyticusintomaize apoplasts6CaMv35s / e1 / nos3′No signal peptide

Abbreviations:

The term “syn-cbh1” refers to a cbh1 gene that has been codon-modified for use in transformation of tobacco plants. It is available from.

The term “CaMV35s” refers to the cauliflower mosaic virus promoter.

The term “SSU” refers to the glycine max rbcS signal...

example 3

[0148] This example shows the construction of plasmids comprising a heterologous gene expression cassette comprising a DNA encoding a ligninase fusion protein and a heterologous gene expression cassette comprising a DNA encoding the bar gene. The constructs are shown in Table 3.

TABLE 3ConstructPlasmid features1rbcSP / ckg4 / pin 3′ / / Act1rbcSP leaf-specificP / bar / nos 3′promoter driving ckg4cDNA of P. chrysosporium2rbcSP / ckg5 / pin 3′ / / Act1rbcSP leaf-specificP / bar / nos 3′promoter driving ckg5cDNA of P. chrysosporium3rbcSP / rbcS SP / ckg4 / pinThe rbcS SP targets ckg43′ / / Act1 P / bar / nos 3′into maize chloroplasts4rbcSP / rbcS SP / ckg5 / pin 3′ / / The rbcS SP targets ckg5Act1 P / bar / nos 3′into maize chloroplasts

Abbreviations:

The terms “ckg4” and “ckg5” mean the ligninase cDNAs isolated from the basidiomycete Phanerochaete. chrysosporium, SEQ ID NO: 11 and SEQ ID NO: 13, respectively. The codons for the 28 amino acid leader are deleted so that the expressed gene product remains inside the cells.

[0149] The ...

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Abstract

The present invention provides transgenic plants expressing one or more cell wall degrading enzymes that can degrade lignocellulose to fermentable sugars. These fermentable sugars can further be fermented to ethanol or other products. The enzymes are directed to the plastids or the apoplasts or the transgenic plant for storage. When the transgenic plants are harvested, the plants are ground to release the enzymes which then are used to degrade the lignocellulose of plant material to produce the fermentable sugars. The transgenic plants express the flowering locus c gene so that flowering is delayed and the plant biomass is increased.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation-in-part of U.S. patent application Ser. No. 11 / 451,162 filed Jun. 12, 2006, which is a continuation-in-part of U.S. patent application Ser. No. 11 / 354,310 filed Feb. 14, 2006, and claims benefit of U.S. patent application Ser. No. 09 / 981,900, filed Oct. 18, 2001, and Provisional Application No. 60 / 242,408 filed Oct. 20, 2000.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT [0002] Not Applicable. REFERENCE TO A “NUCLEOTIDE / AMINO ACID SEQUENCE LISTING APPENDIX SUBMITTED ON A COMPACT DISC”[0003] The application contains nucleotide and amino acid sequences which are identified with SEQ ID NOs. A compact disc is provided which contains the Sequence Listings for the sequences. The Sequence Listing on the compact disc and is identical to the paper copy of the Sequence Listing provided with the application. BACKGROUND OF THE INVENTION [0004] (1) Field of the Invention [0005] The present inventio...

Claims

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Application Information

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IPC IPC(8): A01H5/00C12P19/02C12N15/82
CPCC12N15/8245C12N15/8246C12P19/02C12N15/8261C12N15/827C12N15/8257Y02A40/146
Inventor STICKLEN, MASOMEH B.
Owner BOARD OF TRUSTEES OPERATING MICHIGAN STATE UNIV
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